摘要
利用根癌农杆菌(Agrobacterium tumefaciens)介导,将豇豆胰蛋白酶抑制剂(CpTI)基因导入甘蓝(Brassica oleracea var.capitata L.)栽培品种“京丰”和“迎春”,并分别获得了转基因再生植株。对NPTⅡ的ELISA检测表明,标记基因NPTⅡ已在再生植株中得到有效表达。对转化植株进行Southern blot分子杂交实验进一步证明CpTI基因已整合到甘蓝的基因组。实验室内的叶片离体饲虫及盆栽饲虫试验表明,转基因甘蓝对菜青虫具有一定抗性。实验中还观察到,采用愈伤组织分化途径可以获得较高的转化频率,但嵌合体较多;而直接分芽的途径转化率低,但嵌合体较少。
Cowpea trypsin inhibitor (CpTⅠ) gene was transformed into Brassica oleracea var. capi-tata variety 'Yingchun' and 'Jingfeng' mediated by the Agrobacterium tumefaciens LBA4404 (pRCL27). Transgenic plants were obtained from transformed calli or explants. It was shown from the ELISA assay that NPT Ⅱ gene was expressed in the transgenic cabbage celk. The integration of the CpTⅠ gene into cabbage genome DNA was confirmed by Southern blotting. Insect-tolerance of the transgenic plants to Pieris rapae L. was observed by bioassays on the transgenic plants in the laboratory.
基金
辽宁省科委重点资助项目
关键词
豇豆
胰蛋白酶
抑制剂基因
转基因植株
抗虫
Brassica loeracea var. capitata, Cowpea trypsin inhibitor gene, Agrobacterium tume-faciens, Transgenic plants, Insect-resistance
